(A) Green (530 nm) fluorescence profiles of dissociated testis cells stained with DyeCycle Green, a vital nucleic acid dye. 1C (haploid, spermatids), 2C (diploid, stem cells and secondary spermatocytes) and 4C (tetraploid, primary spermatocytes and mitotic stem cells) populations are marked. Profile for all cells is in orange, 1C 'small' (elongating spermatids, back-gated from plot in B) in green, and 1C 'large' (round spermatids, back-gated from plot in B) in blue. (B) Forward scatter profile for 1C cells gated in A. ‘1C large’ population represents round spermatids. (C) RT-qPCR for sorted round spermatids compared to whole testes. Sycp2 marks spermatocytes, Prm2 marks round and elongating spermatids, Aqp8 marks elongating spermatids only, Lin28a marks spermtogonial stem cells, and Gata4 marks testis somatic cells. Expression is shown relative to whole testis, with Actb as an internal control. Error bars represent SD for three technical replicates. (D) Representatitve 100x images of spermatids stained with DyeCycle Green and sorted directly onto a slide. Scale bars, 5 um.